微卫星DNA分子标记及其在动物遗传育种中的应用

微卫星DNA以其独特的优点和特点而成为当前动物遗传育种研究中颇受欢迎的一种分子标记技术。本文主要介绍了微卫星DNA分子标记的研究简史、原理、流程及其分子标记的优点和在动物育种中的应用。     

分子标记与鸡生产性能的研究进展

随着基因组学时代的到来和各种生物技术的飞速发展 ,分子标记技术在鸡的育种和生产中愈来愈显示其重要性。近几年来 ,国内外学者对微卫星标记进行了广泛而深刻的研究 ,在基因作图、数量性状位点 ( QTL)定位、群体遗传距离测定、抗病品系的选育等方面得到了广泛应用。作为第三代分子标记的 SNPs,为探索鸡的某些疾病的产生机理和生产性能差异的遗传基础提供了新方法。1　微卫星标记微卫星是在生物体基因组以 2～ 6bp为其核心序列 ,头尾相连组成的串状重复序列。核心序列一般重复 1 0～ 2 0次 ,多态性的产生是因重复次数的不同造成的 ,通常…     

RAPD和微卫星分子标记在西方蜜蜂研究中的应用

利用RAPD分子遗传标记 ,构建了西方蜜蜂(Apismellifera)第 1张完整的遗传连锁图 ;确定性基因位点(x)、黑色体色基因位点 (blk)、苹果酸脱氢酶Ⅰ (Mdh - 1)基因位点分别位于第 3、 6、 18个连锁群上 ;确定了影响蜜蜂采粉、报警信息素水平、螫刺行为和体长等数量性状的基因座位 ;可有效地鉴别欧洲蜜蜂和非洲蜜蜂。微卫星技术已成为蜜蜂行为、起源、进化和蜂群内亚家系结构研究的有力工具     

分子标记与地方猪品种的遗传多样性研究

分子标记已成为目前家畜遗传多样性研究的主要工具。本文简要综述了多种分子标记(RFLPs，nat-DNA，RAPD，DNA指纹图谱，微卫星DNA，AFLP，SNPs)的检测方法及其在地方猪品种遗传变异和遗传关系检测分析中的应用，为类似研究提供参考材料。     

微卫星标记及其在家畜遗传育种中的应用

微卫星DNA（Microsatellite DNA）是指以少数几个核苷酸（多数2～4个）为单位多次串连重复的DNA序列, 也称简单重复序列（simple sequence repeat, SSR）、短串联重复（short tandem repeat, STR）或简单序列长度多态性（simple sequence length polymorphism）.早在上世纪70年代初, Skinner在寄居蟹的DNA中发现了一类简单串联重复序列, 1980年Singh等人又在蛇W染色体的性别特意性卫星DNA中发现了一类GATA和GACA的简单重复序列, 1982年Singh等人在人心肌肌动蛋白（Ha-25）的内含子中发现了一个重复25次的（dT-dG）序列,同时指出这一序列在人及其它真核生物的基因组中广泛存在,并与Z-DNA的形成有关.     

ISSR分子标记及其在动物遗传育种中的应用

近年来，分子遗传标记的研究与应用得到了迅速的发展。20世纪80年代中期以来，随着PCR技术的出现，基于PCR技术的分子标记，如随机括增多态性(randomam plified polymorphic DNA，RAPD)、小卫星(minisatellite)或称DNA指纹图谱(DNA fingerprinting，DFP)、微卫星(microsatellite)或称简单序列重复(simple sequence repeat，SSR)、括增片段长度多态性(amplified fragment lengt hpolymorphism，AFLP)、单核     

犬微卫星DNA遗传标记及其在犬繁育中的应用

遗传标记是指可追踪染色体、染色体某一节段、某个基因座在家系中传递的任何一种遗传特性。它具有两个基本特征,一是可遗传性,二是可识别性。因此,生物的任何有差异的突变型均可作为遗传标记。遗传标记主要包括形态标记、细胞学标记、生化标记和DNA分子标记四类,本文对犬微卫星     

DNA分子标记及其在水产动物研究中的应用

分子标记的飞速发展及其广泛的应用,对水产动物的研究工作产生了巨大的影响,在水产动物优良品种的选育、亲缘关系和品系家系的鉴定、重要经济性状基因的定位克隆以及大规模疾病防治等方面发挥重要作用。本文介绍了RFLP,RAPD,AFLP和微卫星DNA等常用分子标记技术的原理和特点,综述了常用分子标记技术在水产动物上的应用,为水产...     

微卫星DNA标记及其在鸡的基因图谱构建中的应用

微卫星ＤＮＡ标记及其在鸡的基因图谱构建中的应用顾志良（江苏省家禽科学研究所扬州２２５００３）王志跃张军（扬州大学农学院扬州２２５００９）１９５３年，Ｗａｔｓｏｎ和Ｃｒｉｃｋ提出的ＤＮＡ分子结构的双螺旋模型圆满地解释了ＤＮＡ就是基因的有机化学实体，宣布...     

微卫星分子标记的研究进展

微卫星是指以少数几个核苷酸(一般1～6个)为单位多次串联重复的DNA序列，是1974年Skinner在研究寄居蟹的卫星DNA时发现的。微卫星广泛均匀地分布在基因组上，其重复数和重复单位序列都是可变的，故多态信息含量大。但由于微卫星无位点特异性，无法确切定位。因此以微卫星核心序列为中心，两侧各加上1个侧翼序列，形成所谓的序列示踪微卫星位点(STMS)。由于侧翼序列在基因组中是单拷贝的，具有位点特异性，而微卫星本身又使STMS具有多态性，只需根据微卫星侧翼序列设计一对PCR引物，通过PCR反应从模板DNA中将该位点扩增出来，然后用聚丙烯酰胺凝胶电泳分离，溴乙锭或银染法显色进行研究。     

利用微卫星分子标记研究我国16份披碱草遗传多样性

利用微卫星(Microsatellite)分子标记对我国16份披碱草Elymus dahuricus进行遗传多样性研究。研究共筛选了38对引物,其中18对引物有扩增条带,占总数的47.4%,具有多态性并且扩增效果较好的引物有6对,占总数的15.8%。用筛选出的这6对引物对供试的16份材料进行微卫星分析,6对引物共检测出26个等位基因,平均每对引物所产生的平均等位基因数为4.3个位点;利用微卫星分子标记数据,对其居群遗传分化进行研究,结果表明:披碱草77.36%的遗传变异出现在居群内,因此,披碱草的遗传多样性主要存在于居群内;对16份披碱草进行聚类分析,构建了亲缘关系树状图,16份供试材料大致分为5类。     

Applying molecular genetic tools to tiger conservation

The utility of molecular genetic approaches in conservation of endangered taxa is now commonly recognized. Over the past decade, conservation genetic analyses based on mitochondrial DNA sequencing and microsatellite genotyping have provided powerful tools to resolve taxonomy uncertainty of tiger subspecies, to define conservation units, to reconstruct phylogeography and demographic history, to examine the genetic ancestry of extinct subspecies, to assess population genetic status non-invasively, and to verify genetic background of captive tigers worldwide. The genetic status of tiger subspecies and populations and implications for developing strategies for the survival of this charismatic species both in situ and ex situ are discussed.     

贵州黎平黄牛线粒体DNA D-loop区全序列初步分析

为了解贵州黎平黄牛的遗传多样性及遗传背景,我们测定了20头黄牛的线粒体DNA D-loop区序列。结果检测到8种单倍型,其中4种为普通牛血统的单倍型,4种为瘤牛血统的单倍型,表明黎平黄牛同时受普通牛和瘤牛的影响。该研究对于黎平黄牛的保种及持续利用有着重要的理论指导意义。     

A lost Sorraia maternal lineage found in the Lusitano horse breed

A common female founder individual of the Portuguese horse breeds Sorraia and Lusitano was found while conducting research on the variation of the Lusitano mitochondrial DNA lineages in relation to studbook information. We obtained 416‐bp control region sequences from 16 descendents of a female Sorraia founder (Pomba) still represented in the living population of the Lusitano, according to the most recent edition of this breed's studbook. The same haplotype was found for all analysed samples and belongs to the haplogroup described by several authors as having predominantly Iberian, South American and North African haplotypes bringing new insights on the relationship between the Sorraia and the other Iberian breeds. This work illustrates how weak the boundary of breed establishment can be, especially at the same geographical region. Using the same founders in different breeds is surely one of the explanations to frequently shared haplotypes among recent breeds, resulting in a lack of consistency between mtDNA sequences and breeds and/or geographical regions.     

Riverine status and genetic structure of Chilika buffalo of eastern India as inferred from cytogenetic and molecular marker-based analysis

The present study aimed at assessing the status of the Chilika buffalo population of eastern India employing cytogenetic and molecular markers. The Chilika buffaloes investigated cytogenetically possess a somatic chromosome count of 50, identical to that of typical riverine buffaloes. Various diversity estimates, viz. observed number of alleles (4.68), effective number of alleles (2.79), and observed (0.487) and expected (0.602) heterozygosity across 25 heterologous microsatellite markers indicated the presence of a moderate level of genetic diversity in Chilika buffaloes, comparable with three other prominent Indian riverine buffalo breeds (Murrah, Nagpuri and Toda) included in this study. Across the four buffalo populations, mean estimates of F -statistics from Jackknifing over loci were significantly different from zero (p < 0.05), with F _IT (total inbreeding estimate) = 0.315 ± 0.038, F _IS (within-population inbreeding estimate) = 0.178 ± 0.038, and F _ST (population differentiation) = 0.166 ± 0.025. Inter-breed analysis reflected Chilika buffaloes to be genetically close to Nagpuri followed by Murrah and Toda buffaloes. Factorial correspondence analysis (FCA) revealed low breed-specific clustering of Chilika and Nagpuri buffaloes. Additionally, the neighbour-joining tree structure of mitochondrial DNA D-loop haplotypes indicated clear grouping of the Chilika haplotypes with the riverine buffalo. Thus the cytogenetic, microsatellite and mitochondrial data analysed in the present study classify Chilika buffalo of eastern India to be of the riverine type and not swamp-type buffalo.     

中国部分山羊品种线粒体DNA D-loop序列遗传多样性分析

分析了中国部分本地山羊品种(18个品种200个体)以及在中国饲养的引入品种(4个品种25个体)线粒体DNA控制区(D-loop)的全序列,结合已报道的世界其它地方的山羊(15个品种77个体)以及2只野山羊的线粒体DNA控制区(D-loop)全序列共304个体,研究结果表明:山羊线粒体DNA控制区约为1 212 bp,检测到228个变异位点,203种单倍型,单倍型多样度为0.993±0.001;核苷酸多样度0.018±0.001。中国部分山羊的变异类型主要为类型A和B,而在巴基斯坦山羊中还检测到类型C和D。比较分析结果表明中国山羊遗传多样性和基因交流比中国黄牛品种要高。     

Characteristics of PCR fragments amplified using five microsatellite markers for identifying the Nagoya breed

The Nagoya breed is a native chicken of Aichi Prefecture, Japan, a dual‐purpose breed for eggs and meat. A method for distinguishing the Nagoya breed from Aichi Prefecture from other chickens using five microsatellite markers (ABR0015, ABR0257, ABR0417, ABR0495 and ADL0262) has already been utilized in order to check the authenticity of Nagoya breed‐labeled chicken on the market. The present study was conducted to investigate nucleotide sequences and sizes of PCR fragments containing the five microsatellite regions for the Nagoya breed and to confirm that the genomic identification could continue to be applied in the future. The DNA sequencing of fragments containing the five markers showed that ABR0015, ABR0417 and ABR0495 had a single haplotype, ABR0257 had three haplotypes, and ADL0262 had two haplotypes, although all the markers exhibited one fixed fragment size each upon sequencing of the fragments and fragment analysis. The results of the fragment analysis of each marker using DNA samples of 28 Nagoya breed males (G0 generation) reared in 2000–2001 and 20 of their offspring males (G8) reared in 2008–2009 showed one fixed fragment size in both populations. Therefore, we confirmed that the five microsatellite markers are useful tools for accurately distinguishing the Nagoya breed from other chickens.     

被动式电子标签在监测高原鼢鼠种群生态变化中的应用

地下啮齿动物种群生态学研究经常需要对动物个体进行标记,而传统的剪耳、剪趾标记法会造成动物伤害、影响动物行为等,与动物福利相悖。利用PIT标签于2014年秋季在祁连山东段标记高原鼢鼠30只,研究高原鼢鼠种群密度及越冬后体重、体长和尾长变化。结果表明,样地内高原鼢鼠估算种群密度为44只/hm2;越冬后高原鼢鼠平均体重为222.07g,显著高于越冬前平均体重193.74g,平均体长为19.4cm,显著高于越冬前平均体长18cm;雌性与雄性的体重增长率无显著性差异(P0.05)。被动式电子标签同时可适用于地下啮齿动物的种群生态学研究,是地下啮齿动物种群生态学研究中的一项新型技术手段。     

贵州威宁黄牛线粒体DNA D-loop区全序列分析

为了解贵州威宁黄牛的遗传多样性及遗传背景，测定了19个个体的线粒体DNAD-loop区全序列。威宁黄牛D-loop区全序列中，A＋T平均含量为61．4％，G＋C含量为38．6％。经比对，共检测到威宁黄牛D-loop区8种单倍型，核苷酸多态位点45个，其中7种为普通牛血统的单倍型，1种为瘤牛血统的单倍型，表明威宁黄牛同时受到普通牛和瘤牛的影响。在威宁黄牛19个个体中，其单倍型多样度为0．715，核苷酸歧异度（π值）为2．415％，表明威宁黄牛品种的遗传多样性丰富。     

A Genetic Diversity Analysis of Mitochondrial DNA D-loop Region in Four High Quality Chicken Breeds

This study was conducted to elucidate the genetic diversity of mitochondrial DNA (mtDNA) D-loop region in Qingyuan partridge chicken group 1,Qingyuan partridge chicken group 2,Yangshan chicken and Qingyuan Yellow feather black-bone chicken.The specific primers were designed according to mtDNA D-loop region of Gullus gullus spadiceus (accession No.:NC_007235.1) in GenBank.The sequence was analyzed after PCR amplification and sequencing,and the haplotype number,polymorphism number,haplotype diversity,nucleotide diversity and nucleotide mean difference were counted.The evolution divergence among breeds was calculated by Mega 5.10 software,and the phylogenetic tree was constructed.The results showed that the length of mtDNA D-loop region in four high quality chicken breeds was 591 bp,and 549 bp were used for subsequent analysis.The content of A,T,C and G were 27.2% to 27.3%,30.1% to 30.4%,29.5% to 29.8% and 12.8% to 12.9%,respectively,and the average content of G+C was 42.5%.There were 92 polymorphic sites which contained 14 singleton variable sites and 78 parsimony informative sites,and the percentage of transitions and transversions were 89.13% (82/92) and 10.87% (10/92),respectively.The haplotype diversity ranged from 0.682 to 0.835,and the nucleotide diversity ranged from 0.00849 to 0.01167.There were 32 haplotypes in all sequences,which could be divided into clades A,B,C and E,however,most of the individuals belonged to clades B (51.2%) and E (37.6%).The phylogenetic tree results showed that four high quality chicken breeds could be classified as 4 branches which were consistent with the haplotypes classification results.The results indicated that the four high quality chicken populations from Qingyuan had relatively high haplotype and nucleotide diversity and likely shared two or more common maternal lineages.